微小RNA-126在子痫前期患者脐血内皮祖细胞中的表达及其意义

Expression and significance of microRNA-126 in endothelial progenitor cells from cord blood in patients with preeclampsia

目的 探讨微小RNA（microRNA,miR）-126在子痫前期发病机制中的作用.方法 选择2011年11月至2012年4月在华中科技大学同济医学院附属协和医院妇产科剖宫产分娩的正常妊娠孕妇20例、子痫前期孕妇24例,于剖宫产术中采集脐静脉血,分离并培养内皮祖细胞（endothelial progenitor cells,EPCs）.采用逆转录-聚合酶链反应技术检测EPCs中miR-126的表达水平.将正常妊娠孕妇的EPCs分为实验组、阴性对照组及空白对照组.实验组转染miR-126mimics,阴性对照组转染无意义的阴性对照小干扰RNA寡核苷酸,空白对照组不转染.采用逆转录-聚合酶链反应技术检测转染后EPCs中miR-126的表达,显微镜下计数梭形细胞（即处于分化期的EPCs）个数,采用Transwell模型迁移实验检测迁移细胞数,采用小管形成实验观察管腔形成数.采用两独立样本t检验、方差分析及q检验进行统计学分析. 结果 子痫前期孕妇脐血EPCs中miR-126的相对表达水平低于正常妊娠孕妇[（37.50±14.81）％与（99.00±2.00）％,t=9.40,P＜0.05].转染后实验组EPCs中miR-126的表达水平为（735.50±40.35）％,梭形细胞数、迁移细胞数和管腔形成数分别为（130.00±3.61）、（128.33±7.51）和（59.33±6.66）个,均高于阴性对照组[分别为（98.31±5.25）％,（53.45±3.00）、（70.67±4.71）和（25.67±3.51）个,q值分别为76.00、42.89、17.75和18.24,P值均＜0.01],也高于空白对照组[分别为（100.00±0.00）％,（59.33±2.51）、（83.34±4.21）和（29.35±4.34）个,q值分别为75.70、39.60、13.84和16.25,P值均＜0.01];但空白对照组和阴性对照组比较差异均无统计学意义（q值分别为0.24、3.29、2.87和1.98,P值均＞0.05）. 结论 EPCs miR-126高表达可能通过促进EPCs的分化、迁移和管腔形成能力,增强血管形成和修复;EPCs miR-126低表达可能与子痫前期的发生有关.

Objective To investigate the role of microRNA （miR）-126 in the pathogenesis of preeclampsia.Methods From November 2011 to April 2012,20 healthy pregnant women and 24 pregnant women with preeclampsia underwent cesarean section in Union Hospital of Tongji Medical College were selected.Umbilical venous blood was taken and the endothelial progenitor cells （EPCs） were separated and cultured.Reverse transcription-polymerase chain reaction （RT-PCR） was used to test the level of miR-126 in cultured EPCs.EPCs separated from healthy pregnant women were divided into the study group,negative group and control group,and transfected with miR-126 mimics,negative small interfering RNA and no small interfering sequence,respectively.RT-PCR was used to measure the levels of miR-126 of EPCs,and spindle cells （at differentiation stage of EPCs） were counted under microscope.Changes in migration and tube formation capacity of EPCs were detected by Transwell and tube formation test,respectively.Two sample t test,analysis of variance and q test were used for statistical analysis.Results Expression ofmiR-126 in preeclampsia pregnant women was decreased compared with that in healthy pregnant women [（37.50± 14.81） ％ vs （99.00±2.0） ％,t=9.40,P＜0.05].In the study group,the expression of miR-126 was （735.50±40.35） ％,and the number of spindle cells,migration cells and branch points was 130.00±3.61,128.33 ±7.51 and 59.33 ±6.66.These were higher than in the negative group [（98.31 ±5.25） ％,53.45±3.00,70.67±4.71 and 25.67±3.51; q=76.00,42.89,17.75 and 18.24,all P＜0.01],and the control group [（100.00±0.00） ％,59.33±2.51,83.34±4.21 and 29.35±4.34; q=75.70,39.60,13.84 and 16.25,all P＜0.01].However,there were no significant differences between the negative and control groups （q=0.24,3.29,2.87 and 1.98,all P＞0.05）.Conclusions High expression of miR-126 could improve the proliferation,migration and tube formation capacity of EPCs.Low expression of miR-126 in pregnant women may be involved in the pathogenesis of preeclampsia.